澳洲茄胺对人骨肉瘤移植瘤的抑制作用及其机制  

Inhibitory effect of solasodine on human osteosarcoma xenografts and its mechanism

作　　者：杨利[1] 李小健[1] 李朝军[1] YANG Li;LI Xiaojian;LI Chaojun(Department of Orthopaedics,Second People's Hospital of Tangshan Fengrun District,Tangshan,Hebei 064000,China)

机构地区：[1]唐山市丰润区第二人民医院骨科,河北唐山064000

出　　处：《安徽医药》2021年第4期664-668,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨澳洲茄胺对人骨肉瘤移植瘤的抑制作用及其机制。方法该研究于2018年1月至2019年3月完成。MG63细胞株购于中科院上海细胞库。将0.2 mL人骨肉瘤MG63细胞悬液(共2×106个细胞)注射于裸鼠右侧腋窝皮下,建立人骨肉瘤移植瘤模型,实验分为4组,即对照组和澳洲茄胺组(10 mg/kg、20 mg/kg、40 mg/kg),隔日腹腔注射给药1次,连续4周。测量移植瘤的质量和体积,原位末端标记(TUNEL)染色法检测细胞的凋亡,蛋白质印迹法检测凋亡相关蛋白、血管生成相关因子和磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路的表达。结果与对照组相比,澳洲茄胺组的移植瘤质量(0.45±0.12)g和在21 d(510.62±95.78)mm~3和28 d(579.56±92.04)mm~3的体积降低,且以40 mg/kg组的抑制效应最明显(P<0.05)。与对照组相比,澳洲茄胺组的原位末端标记的异硫氰酸荧光素(TUNEL-FITC)的荧光强度升高,且以40 mg/kg组最明显。与对照组相比,澳洲茄胺组移植瘤中B细胞淋巴瘤/白血病-2相关X蛋白(Bax)(3.55±0.39)、活化的半胱氨酸天冬氨酸蛋白酶-3(Cleaved caspase-3)(3.94±0.33)、活化的半胱氨酸天冬氨酸蛋白酶-9(Cleaved caspase-9)(3.59±0.31)、凝血酶敏感蛋白1(TSP-1)(3.70±0.41)蛋白的表达量上调,B细胞淋巴瘤/白血病-2(Bcl-2)(0.31±0.15)、血管内皮生长因子(VEGF)(0.25±0.12)、磷酸化磷脂酰肌醇3-激酶(p-PI3K)(0.39±0.12)和磷酸化蛋白激酶B(p-AKT)(0.33±0.11)蛋白的表达量下调,且以40 mg/kg组的效应最明显(P<0.05)。结论澳洲茄胺可以抑制人骨肉瘤细胞皮下移植瘤的生长,诱导细胞凋亡,下调VEGF的表达,上调TSP-1的表达,其作用机制可能与PI3K/AKT信号通路的抑制有关。Objective To explore the inhibitory effect of solasodine on human osteosarcoma xenografts and its mechanism.Meth⁃ods The study was completed from January 2018 to March 2019.The MG63 cell line was purchased from Shanghai Cell Bank of Chinese Academy of Sciences.The human osteosarcoma MG63 cell suspension(0.2 mL,2×106 cells)was injected subcutaneously into the right armpit of nude mice to establish human osteosarcoma xenograft model.The experiment was assigned into four groups:control group and solasodine group(10 mg/kg,20 mg/kg,40 mg/kg).The mice were injected intraperitoneally once every other day for 4 weeks.The weight and volume of xenografts were measured.Apoptosis was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The expression of apoptosis-related proteins,angiogenesis-related factors and phosphoinositide-3-kinase/serine-threonine kinase(PI3K/AKT)signaling pathway were detected by Western blotting.Results Compared with control group,the weight(0.45±0.12)g and volume(510.62±95.78)mm3 at 21 d and(579.56±92.04)mm3 at 28 d of the xenografts in the solasodine group decreased,and the inhibitory effect of 40 mg/kg group was the most obvious(P<0.05).The fluorescence intensity of terminal-deoxynucleotidyl transferase mediated nick end labeling-fluorescein-isothiocyanate(TUNEL-FITC)in the solasodine group was higher than that in the control group,especially in the 40 mg/kg group.The protein expression of B cell lymphoma/leukemia-2 associated X protein(Bax)(3.55±0.39),cleaved cystein-asparate protease-3(Cleaved caspase-3)(3.94±0.33),cleaved cystein-asparate protease-9(Cleaved caspase-9)(3.59±0.31),thrombospondin 1(TSP-1)(3.70±0.41)were up-regulated,while the protein expression of B cell lymphoma/leukemia-2(Bcl-2)(0.31±0.15),vascular endothelial growth factor(VEGF)(0.25±0.12),phosphorylated phosphoinositide-3-kinase(p-PI3K)(0.39±0.12)and phosphorylated serine-threonine kinase(p-AKT)(0.33±0.11)were down-regulated,and the effect of 40 mg/kg group was the most obv

关 键 词：澳洲茄胺 龙葵 人骨肉瘤细胞 细胞凋亡 血管内皮生长因子 凝血酶敏感蛋白1 

分 类 号：R285[医药卫生—中药学]